ABSTRACT: DNA samples of haploid ES cells and control DNA were compared to genomic DNA of the C57B/6 inbred mouse strain on a NimbleGen Mouse CGH 3x720K Whole-Genome Tiling Array (Build MM9), (GPL10989) DNA samples of haploid ES cells and control DNA were compared to genomic DNA of the 129/Sv inbred mouse strain on a NimbleGen Mouse CGH 3x720K Whole-Genome Tiling Array (Build MM9), (GPL10989)
Project description:DNA samples of haploid ES cells and control DNA were compared to genomic DNA of the C57B/6 inbred mouse strain on a NimbleGen Mouse CGH 3x720K Whole-Genome Tiling Array (Build MM9), (GPL10989) this analysis was performed to analyse copy number variations in haploid mouse ES cells from different genetic backgrounds. Male C57/B6 DNA was used as control. To identify CNVs based on the different genetic background control experiments comparing DNA from the inbred CBA and a mixed transgeneic mouse strain were performed.
Project description:Comparative genomic hybridization (CGH) analysis of androgenetic haploid embryonic stem cells was done with 129/Sv male mouse kidney genomic DNA as reference. The results revealed no consistent copy number variations (CNVs) among the examined cell lines (AH129-5, AH129-N1 and AH129-NC1), except for the global difference on the X chromosome to the male control. Genomic DNA of FACS sorted G0/G1 stage ahES cells was compared with 129/Sv male mouse kidney genomic DNA reference by the NimbleGen Mouse CGH 3x720K Whole-Genome Tiling Array (Build MM9) [100718_MM9_WG_CGH_HX3]
Project description:The genomic DNA sample of AG-haESCs were compared to the C57BL/6J male mouse kidney by comparative genomic hybridization. The data confirmed that the haploid cells sustained genome integrity. The analysis was performed on a NimbleGen Mouse CGH 3x720K Whole-Genome Tiling Array to analyse the copy number variations in AG-haESCs, and the genomic DNA of C57BL/6J male mouse kidney was used as control, which had the same background with haploid ESCs.
Project description:Comparative genomic hybridization (CGH) analysis of CiPSCs (clone CiPS-57 and CiPS-58) and OSKM-iPSCs. C57BL/6 mouse embryonic fibroblasts (MEFs) genome DNA as a reference. No consistent copy number variations (CNVs) among CiPSCs and OSKM-iPSCs. Genome DNA of CiPSCs and OSKM-iPSCs (both of them induced form C57BL/6 MEFs) was isolated and compared with C57BL/6 MEFs genomic DNA reference by the NimbleGen Mouse CGH 3x720K Whole-Genome Tiling Array.
Project description:Testicular germ cell tumors are among the most responsive solid cancers to conventional chemotherapy. To elucidate the underlying mechanisms, we developed a mouse testicular germ cell tumor model in which germ cell-specific oncogenic Kras activation and tumor suppressor Pten inactivation was driven by CRE-mediated recombination. The resulting mice rapidly developed malignant, metastatic testicular cancers composed of both teratoma and embryonal carcinoma, the latter of which exhibited stem cell characteristics, including expression of the pluripotency factor OCT4. As part of our analysis of mouse gPAK testicular tumors, as well as comparison to benign 129-Dnd1Ter/Ter testicular teratomas, we used NimbleGen Mouse CGH 3x720k Whole-Genome Tiling Arrays to assess copy number variations in this novel genetically engineered mouse model of malignant, metastatic testicular cancer.
Project description:gDNA was obtained from 26 frozen ERMS tumors for aCGH and copy number analysis Tumors compared to nimblegen pooled male reference, using 2.1 Million probe Nimblegen array CGH Build 18 NCBI36
Project description:Breast cancer is the most prevalent cancer in women 1, and most cases are believed to have a sporadic, rather than heritable basis 2. To identify breast cancer driver genes, we exploited the C3H-Mcm4Chaos3/Chaos3 (“Chaos3”) mouse model that, by virtue of bearing a defective DNA replicative helicase subunit that causes elevated genomic instability (GIN), sustains somatic alterations ultimately causing mammary adenocarcinomas 6. Array Comparative Genomic Hybridization (aCGH) analysis of Chaos3 mammary tumors revealed recurrent copy number alterations (CNAs), most notably deletion of the Neurofibromin 1 (Nf1) tumor suppressor gene in all cases. NF1, a negative regulator of RAS, is traditionally recognized for its role in driving the development of neurofibromas in the context of the human disease Neurofibromatosis Type 1, but not breast cancer. Genomic DNA from tumor and reference samples were hybridized to NimbleGen 3x720K mouse CGH arrays. Two reference samples were used independently. CNAs were visualized using Nimblegen, IGV, and KCsmart software 32. Select genes were validated via qPCR. Critical regions within each Chaos3 CNA were identified as the region with the greatest overlap across multiple Chaos3 tumors.
Project description:Acute Promyelocytic Leukemia (APL) is characterized by the t(15;17)(q22;q11.2) translocation, which creates a PML-RARA fusion gene that can initiate APL in mice. To discover cooperating mutations in this model, we sequenced a mouse APL genome to 15.6x haploid coverage, and discovered three somatic, non-synonymous mutations, of which one (Jak1 V657F) was recurrent. This mutation is identical to the JAK1 V658F mutation previously found in human APL and ALL samples. JAK1 V658F cooperates in vivo with PML-RARA, causing a rapidly fatal leukemia. We also discovered a somatic 150kb deletion involving the Kdm6a/Utx gene in the mouse APL genome; 3/14 additional mouse APL samples had similar deletions involving Kdm6a/Utx. Kdm6A/Utx, a histone H3K27 demethylase, was also deleted in 1/150 human AML samples tested. Whole genome sequencing of mouse cancer genomes provides an unbiased approach for discovering functionally relevant mutations that are also present in human leukemias. DNA from 15 mouse APL tumors on the Bl/6 Taconic background (F10), DNA from one WT 129/SvJ mouse spleen, and pooled DNA derived from the spleens of 6-week-old, wild type, Bl/6 Taconic (parental strain) mice were analyzed using the Nimblegen Mouse CGH 3x720K WGT platform.
Project description:This study used the NimbleGen dog whole genome CGH 2.1M tiling array to assay copy number variants in the dog genome in multiple breeds and wolf.
Project description:It is well-known that embryonic stem cells (ESC) are much more sensitive to replication-induced stress than differentiated cells but the underpinning mechanisms are largely unknown. H2A.X, a minor variant of H2A, constitutes only 1-10% of the mammalian genome. H2A.X plays a well-known for role in the DNA damage response and maintaining stability in the genome, including the regions frequently experiencing replication stress, such as the fragile sites. Intriguingly, several recent studies have reported that H2A.X function is elevated in ESC; and others reported that H2A.X function is provoked during cellular reprogramming (in induced pluripotent stem cells, iPSC), indicating that increased proliferation during iPS may trigger replication stress and the H2A.X DNA damage response. However, several studies of genomic instability in iPSC led to different conclusions on this important issue. For example, frequent copy number variants (CNV) were reported at the genomic regions sensitive to replication stress, such as the fragile sites. On the other hand, another study reported the lack of genomic instability in mouse iPS clones that are able to generate “all-iPS” animals in tetraploid complementation assays (4N+ iPSC), indicative of a potential link between pluripotency and genome integrity. However, whether if high level genomic instability occurs in the 4N- iPSC iPSC clones at replication stress sensitive regions is unknown. Moreover, due to the lack of mechanistic insights on genome integrity maintenance, how pluripotency and genome integrity are connected remains elusive. Here we show that H2A.X plays unexpected roles in maintaining pluripotency and genome integrity in ESC and iPSC. In ESC, it is specially enriched at genomic regions sensitive to replication stress so that it protects genome integrity thereat. Faithful H2A.X deposition is critical for genome integrity and pluripotency in iPSC. H2A.X depositions in 4N+ iPSC clones faithfully recapitulate the ESC pattern and therefore, prevent genome instability. On the other hand, insufficient H2A.X depositions in 4N- iPSC clones at such regions lead to genome instability and defects in replication stress response and DNA repair, reminiscent of the H2A.X deficient ESC. In this study, male 129sv/C57 ES cell genomic DNA was used as reference control, to identify CNV sites in iPS cell lines. And also detect H2A.X (-/-) ES cell (129/Sv) CNVs, with the H2A.X(f/f) ES cell DNA (129/Sv) as control. DNA samples were compared on NimbleGen Mouse CGH 3x720K Whole-Genome Tiling Array (Build MM9).